The Use of Aptamers in Breast Cancer Anti-Metastasis Therapy
Abstract
The objective of this one-year proposal was to initiate studies directed to identify and generate oligonucleotides (aptamers) that block the adhesion between tumor cell beta 4 integrin and endothelial hCLCA2, an adhesion interaction implicated in pulmonary vascular arrest and metastasis of human breast cancer cells. Phase one of these studies was concerned with the generation of an 'aptamer bait' for SELEX random sequence ssRNA library screening. Baits consisted of wild-type, full length beta 4 protein, a polypeptide comprising the beta 4 amino acids 108 to 343 (I-domain like structure), and a synthetic beta 4 integrin-peptide representing the hCLCA2-binding domain. The first two 'baits' proved to be useless due to poor harvest, insufficient purity, and loss of stability in case of full-length beta 4 or insolubility of the 108-343 polypeptide prepared in insect cells. Identification of the hCLCA2-binding domain of beta 4 was accomplished by domain swapping between the beta 1 and beta 4 integrins within the I-domain-like structure. A sequence of 20 amino acids was identified within that structure that harbors the hCLCA2-binding site. This sequence was synthesized and found to effectively block the beta 4 integrin/hCLCA2 adhesion. Selection of aptamers is in progress using this promising, reproducible bait.
Document Details
- Document Type
- Technical Report
- Publication Date
- Sep 01, 2001
- Accession Number
- ADB278762
Entities
People
- Besdicht U. Pauli
Organizations
- Cornell University